Fine Tuning Antibody Conjugation Methods using SNAP-tag Technology

Chouman, Karinna; Woitok, Mira; Mladenov, Radoslav; Kessler, Claudia; Weinhold, Elmar G.; Hanz, Gisela Maria; Fischer, Rainer; Meinhold‐Heerlein, Ivo; Bleilevens, Andreas; Gresch, Gerrit; Haugg, Anka Maria; Zeppernick, Felix; Bauerschlag, Dirk; Maass, Nicolaì; Stickeler, Elmar; Kolberg, Katharina; Hussain, Ahmad Fawzi

doi:10.2174/1871520617666170213123737

Cited by 3 publications

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“…The toxicity of the 647-CLIP-scFv-425-SNAP-IRDye700 and scFv-425-SNAP-IRDye700 proteins against EGFR + ovarian cancer cell lines (OVCAR-3, SKOV-3, and IGROV-1) and the EGFR – cell line (A2780) was determined using the XTT II cell proliferation kit (Roche, Mannheim, Germany) as previously described. ,, All experiments were performed at least three times in triplicates, and the data are presented as means ± standard error of the mean (SEM). The reagent concentration required to eliminate 50% of the ovarian cancer cells (IC 50 ) relative to control cells was determined using GraphPad Prism v 5 (GraphPad, San Diego, CA).…”

Section: Methodsmentioning

confidence: 99%

Simultaneous and Independent Dual Site-Specific Self-Labeling of Recombinant Antibodies

et al. 2018

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Antibody-based diagnostic and therapeutic reagents armed with effector molecules such as dyes and drugs offer hope in the battle against cancer. Several site-specific conjugation methods have been developed to equip antibodies with such effector molecules, but they tend to be expensive and involve multiple reaction steps. The conjugation of two different effector molecules to a single antibody also remains a major challenge. Here we describe a simple, controlled, and robust method for the dual site-specific conjugation of an antibody with two effector molecules in a single-pot reaction using the selflabeling SNAP and CLIP protein tags. We verified the principle of the method by labeling an epidermal growth factor receptor (EGFR)-specific single-chain antibody fragment (scFv-425) simultaneously with IRDye700 and Alexa-Fluor647. This dual-labeled antibody bound to EGFR + ovarian cancer cell lines and tissue samples with high specificity, and its phototherapeutic efficacy was confirmed by the selective killing of EGFR + cells in vitro.

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Section: Methodsmentioning

confidence: 99%

Simultaneous and Independent Dual Site-Specific Self-Labeling of Recombinant Antibodies

et al. 2018

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“…Conjugating the AURIF to anti-Her2 and anti-EGFR recombinant antibodies significantly improved the specificity of AURIF and increased its cytotoxicity by more than 30-fold toward Her2 expressing breast cancer cells [46]. Furthermore, the highly potent near-infrared (NIR) imaging/photosensitizer IRDye700 (phototheranostics) agent was conjugated to several recombinant antibody fragments targeting different cancer cell-surface receptors using SNAP-tag technology to generate a panel of NIR photoimmunotheranostics (NIR-PIT) agents [47,81,[84][85][86].…”

Section: O6-alkylguanine-dna Alkyltransferase (Agt) (Snap-tag)mentioning

confidence: 99%

Toward Homogenous Antibody Drug Conjugates Using Enzyme-Based Conjugation Approaches

et al. 2021

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In the last few decades, antibody-based diagnostic and therapeutic applications have been well established in medicine and have revolutionized cancer managements by improving tumor detection and treatment. Antibodies are unique medical elements due to their powerful properties of being able to recognize specific antigens and their therapeutic mechanisms such as blocking specific pathways, antibody-dependent cellular cytotoxicity, and complement-dependent cytotoxicity. Furthermore, modification techniques have paved the way for improving antibody properties and to develop new classes of antibody-conjugate-based diagnostic and therapeutic agents. These techniques allow arming antibodies with various effector molecules. However, these techniques are utilizing the most frequently used amino acid residues for bioconjugation, such as cysteine and lysine. These bioconjugation approaches generate heterogeneous products with different functional and safety profiles. This is mainly due to the abundance of lysine and cysteine side chains. To overcome these limitations, different site-direct conjugation methods have been applied to arm the antibodies with therapeutic or diagnostics molecules to generate unified antibody conjugates with tailored properties. This review summarizes some of the enzyme-based site-specific conjugation approaches.

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Near Infrared Photoimmunotherapy: A Review of Recent Progress and Their Target Molecules for Cancer Therapy

Mohiuddin

Zhang

Sheng

et al. 2023

IJMS

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Near infrared photoimmunotherapy (NIR-PIT) is a newly developed molecular targeted cancer treatment, which selectively kills cancer cells or immune-regulatory cells and induces therapeutic host immune responses by administrating a cancer targeting moiety conjugated with IRdye700. The local exposure to near-infrared (NIR) light causes a photo-induced ligand release reaction, which causes damage to the target cell, resulting in immunogenic cell death (ICD) with little or no side effect to the surrounding normal cells. Moreover, NIR-PIT can generate an immune response in distant metastases and inhibit further cancer attack by combing cancer cells targeting NIR-PIT and immune regulatory cells targeting NIR-PIT or other cancer treatment modalities. Several recent improvements in NIR-PIT have been explored such as catheter-driven NIR light delivery, real-time monitoring of cancer, and the development of new target molecule, leading to NIR-PIT being considered as a promising cancer therapy. In this review, we discuss the progress of NIR-PIT, their mechanism and design strategies for cancer treatment. Furthermore, the overall possible targeting molecules for NIR-PIT with their application for cancer treatment are briefly summarised.

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Fine Tuning Antibody Conjugation Methods using SNAP-tag Technology

Abstract: Here, we describe a simple, rapid and robust site-directed labeling method that can be used to generate homogeneous immunoconjugate with defined pharmacological properties.

Cited by 3 publications

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Simultaneous and Independent Dual Site-Specific Self-Labeling of Recombinant Antibodies

Simultaneous and Independent Dual Site-Specific Self-Labeling of Recombinant Antibodies

Toward Homogenous Antibody Drug Conjugates Using Enzyme-Based Conjugation Approaches

Near Infrared Photoimmunotherapy: A Review of Recent Progress and Their Target Molecules for Cancer Therapy

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